Capillary (Sanger) DNA Sequencing

The GGBC maintains an Applied Biosystems 3730xl 96-capillary DNA Analyzer for Sanger sequencing and fragment analysis. Users can submit 96 well plates of products that are ready to be sequenced or choose to do the cycle sequencing reaction themselves and we will process the “Ready-to-Run” plates on the 3730xl. Individual customer sequence read length will vary.

Customers may download data directly from their FBS Portal account. Upon request, data may be delivered via SFTP. Users are assigned password protected folders. All data resides within the PI’s folder, however independent subfolders can be obtained if requested.

Sample Preparation

Plate submissions for Sanger Sequencing

Please combine template DNA and primer using the following guidelines:

Template quantity required for sequencing

Purified Plasmid or Purified PCR Product (bp)	Quantity Required (total ng)
100-200	1-3
200-500	3-10
500-1000	5-20
1000-2000	10-40
>2000	20-50
Double-stranded Plasmid	150-300
*Cosmid, BAC	*0.5-1.0 µg

Sample volume should be between 2-10µL

Sequencing Primers

Primer concentrations 3.3µM

Per each reaction, use 1 µL primer (at 3.3µM)

Ready-to-run sequencing

Customers who want to do the cycle sequencing themselves can submit reactions in 96-well plates. Reactions should be submitted wet in high quality molecular grade water (if submitted dry, please let us know how much water to rehydrate them in). The reactions have to be in semi-skirted plates that are compatible with the 3730xl.

Please submit samples in a 96-well plate as GGBC no longer accepts single tube submissions. Please submit ALL samples in a SEMI-SKIRTED 96-well plate.

PLEASE USE CONICAL BOTTOM PLATES AND NOT ROUND/FLAT BOTTOM PLATES.

Submit Samples

UGA and on-campus customers:

Please go to the FBS Portal page to submit your order, and then bring your samples to the GGBC at 110 Riverbend Rd., Athens, GA 30602 (Room 156) anytime Monday-Friday between 8:00 am-5:00 pm.

Please do not tape your samples to your order sheet.

Off-campus customers:

Please ship samples to this address:

University of Georgia
Georgia Genomics and Bioinformatics Core
110 Riverbend Rd, Rm 161
Athens, GA 30602

Tips: Please seal plates with plate sealing foil and ship on dry-ice or in freezer packs preferably through FedEx or UPS as they deliver directly to GGBC. You might also want to write “this side up” on your cooler or box.

Data Retrieval

Please go to the FBS Portal page to retrieve your data.

Prices and Quotes

Contact for Financial Inquiries and Quote Requests

Please email Kim and Elizabeth at ggbc@uga.edu, for financial inquiries or to request a quote. Be as specific as possible, so that they can more quickly assist you.

Current Price List

Service Description	UGA Fee	Non-UGA Fee	Commercial Fee
Ready to run genotyping plate	$75.00	$88.50	$93.75
Ready to run sequencing plate	$75.00	$88.50	$93.75
Sequencing a 96 well plate	$260.00	$307.00	$325.00
Sephadex clean up/per plate	$50.40	$59.50	$63.00
Sephadex clean up/per half plate	$29.00	$34.25	$36.25
Semi-skirted 96-well plate (for 3730 sequencer)	$19.00	$22.45	$23.75
Normalize DNA concentration in a 96 well plate	$86.00	$102.00	$108.00

Prices are subject to change without notice. To get a quote, please contact GGBC.

Data Analysis

Any of the following programs can be used to view DNA traces, but training and support are not available through GGBC .

Low-cost research data storage is available through the Georgia Advanced Computing Resource Center.

Capillary Sequencing Protocols

The following files are available for download in PDF format: